CN110272808B - Nucleic acid extraction system - Google Patents
Nucleic acid extraction system Download PDFInfo
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- CN110272808B CN110272808B CN201810206072.XA CN201810206072A CN110272808B CN 110272808 B CN110272808 B CN 110272808B CN 201810206072 A CN201810206072 A CN 201810206072A CN 110272808 B CN110272808 B CN 110272808B
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Abstract
The invention discloses a nucleic acid extraction system, and belongs to the field of experimental instruments. The nucleic acid extraction system includes: nucleic acid extraction board and the pipetting device and the positive pressure device that set up side by side, the pipetting device includes: transfusion system, transfusion system mounting bracket, transfusion system drive unit and a plurality of infusion unit, every the infusion unit all includes: a reagent supply and a reagent flow controller; the positive pressure device includes: the device comprises a plug, a vent pipe, a positive pressure provider and a positive pressure driving device. The nucleic acid extraction system has high efficiency and low cost when extracting nucleic acid.
Description
Technical Field
The invention relates to the field of experimental instruments, in particular to a nucleic acid extraction system.
Background
When the magnetic bead method or the column method is used for automatically extracting or purifying nucleic acid, at least one of an automatic pipetting workstation, a centrifuge and a negative pressure machine is needed. The existing nucleic acid extraction or purification process comprises: cell lysis, nucleic acid adsorption, nucleic acid washing and nucleic acid elution. In the process, 5-6 chemical reagents for extracting or purifying nucleic acid are transferred to a centrifuge tube through an automatic liquid transfer workstation by means of a liquid transfer sucker to crack cells or to an adsorption column to clean and elute the nucleic acid, the centrifuge tube and the adsorption column are transferred to a centrifuge through the automatic liquid transfer workstation, solid phase impurities in the centrifuge tube are precipitated through the centrifugal action, and liquid in the adsorption column flows downwards through the resistance of a membrane through the centrifuge or a negative pressure machine to achieve the effect of cleaning and eluting the nucleic acid on the membrane.
In the process of implementing the invention, the inventor finds that the prior art has at least the following problems:
when existing nucleic acid is extracted or purified, the action is slow, the time consumption is long and the cost is high when an automatic pipetting workstation is used for pipetting, for example, the process of transferring 1 solution each time is as follows: after the liquid transfer suction head is accurately aligned, the liquid transfer suction head is lifted, moved to a reagent basin for liquid absorption, moved to an upper opening of each hole on a centrifuge tube or an adsorption column or a 96-hole plate for releasing reagent, continuously absorbs and releases liquid so as to uniformly mix the liquid, and moved to a garbage can for discarding the suction head. If 6 reagents for extracting or purifying nucleic acid are to be transferred longer than necessary at that time. In addition, when pipetting, not only the number of pipette tips is large and the quality requirement is high, otherwise the tips cannot be inserted into the extraction and pipetting accurately, which increases the pipetting cost, and generally, when extracting and purifying nucleic acid with high throughput, at least 6 solutions are transferred into a 96-well plate, which requires 6 cassettes of tips with high quality, so that the pipetting method is inefficient and costly when extracting or purifying nucleic acid. Meanwhile, when the centrifuge is used, the centrifuge tube and the adsorption column need to be repeatedly placed into the centrifuge for centrifugation and then taken out after centrifugation, and the repeated operation process is complicated and consumes long time. In order to reduce the use of the centrifugal machine, a negative pressure machine can also be adopted, specifically, the adsorption column is inserted into a negative pressure box connected with the negative pressure machine, and the cleaning liquid in the adsorption column flows into a waste liquid basin in the negative pressure box through negative pressure. However, when a negative pressure machine is used, the suction force of the negative pressure machine is reduced due to the blocking of the waste liquid basin, so that the negative pressure provided by the negative pressure machine is not strong enough, and the negative pressure generated by the negative pressure machine used for experiments at present is not strong enough, so that the negative pressure machine can only be used in the step of cleaning in the nucleic acid extraction process, but not in the whole process, and the method for extracting or purifying nucleic acid still has the problems of complicated process, long time consumption and low efficiency.
Disclosure of Invention
In order to solve the problems of low working efficiency and high cost when an automatic liquid transfer workstation is matched with a centrifuge or a negative pressure machine to extract nucleic acid in the prior art, the embodiment of the invention provides a nucleic acid extraction system. The technical scheme is as follows:
the embodiment of the invention provides a nucleic acid extraction system, which comprises: a mounting rack, a nucleic acid extracting plate, a liquid-transfering device and a positive pressure device which are arranged side by side,
the liquid transfer device includes: the infusion set comprises sealing cavities which correspond to the infusion units one by one, the sealing cavities are arranged side by side, a needle is arranged at the bottom of each sealing cavity, each needle is communicated with the corresponding sealing cavity, the infusion set is arranged at the bottom of the infusion set mounting frame, the infusion set driving unit is fixed on the mounting frame and is configured to drive the infusion set to move up and down through the infusion set mounting frame;
each of the infusion units comprises: a reagent supply and a reagent flow controller, wherein for any one of the infusion units, a liquid outlet of the reagent supply is communicated with a liquid inlet of the reagent flow controller, and a liquid outlet of the reagent flow controller is communicated with each sealed cavity;
the positive pressure device includes: the air supply device comprises a plug, an air pipe, a positive pressure provider and a positive pressure driving device, wherein the plug is provided with an air pipe communication hole, one end of the air pipe is in sealed communication with the air pipe communication hole, the other end of the air pipe is in sealed communication with an air outlet of the positive pressure provider, the positive pressure driving device is fixed on the mounting frame and is configured to drive the positive pressure provider to move up and down;
be provided with a plurality of nucleic acid on the nucleic acid extraction board and draw the post, a plurality of nucleic acid are drawn the post and are arranged correspondingly syringe needle or the below of end cap, every all be provided with on the inlet of nucleic acid extraction post with the sealed complex sealed pad of end cap, nucleic acid draw the post with correspond sealed connection can be dismantled to the end cap.
Specifically, the nucleic acid extraction system further comprises: mixing board, mixing board mounting bracket and mixing board drive arrangement, mixing board mounting bracket is fixed on the mounting bracket, mixing board drive arrangement drive is installed on the mixing board mounting bracket, the mixing board is installed the bottom of mixing board mounting bracket, the bottom of mixing board be provided with the mixing stick of a plurality of nucleic acid extraction post one-to-one, mixing board drive arrangement is configured to pass through mixing board mounting bracket drive the mixing stick is in reciprocate in the nucleic acid extraction post.
Further, the nucleic acid extraction system further comprises: the first partition plate is arranged between the mixing rod and the nucleic acid extraction plate, the first partition plate driving device is fixed on the mounting frame, and the first partition plate driving device is configured to drive the first partition plate to horizontally move below the mixing rod.
Further, the nucleic acid extraction column includes: a filter column comprising a breakable member and a plurality of filter members, the breakable member being disposed between a liquid inlet of the nucleic acid extraction column and a liquid outlet of the nucleic acid extraction column, and is fixed on the nucleic acid purification column, a groove is arranged on the breakable part, the bottom of the mixing rod is a conical structure for poking the groove, the mixing rod is sleeved with a mixing sheet, the outer diameter of the mixing sheet is smaller than the inner diameter of the nucleic acid extraction column, the plurality of filter members are disposed between the breakable member and the liquid outlet of the nucleic acid extraction column, and are respectively fixed on the inner wall of the nucleic acid extraction column, each filter element is provided with a plurality of filter micropores, and two adjacent filter elements are arranged in the filter element, the pore diameter of the filtering micropores of the filtering piece close to the liquid inlet of the filtering column is larger than that of the filtering micropores of the filtering piece close to the liquid outlet of the filtering column.
Further, the nucleic acid extraction column includes: adsorption column, adsorption column is including brokenly the part and adsorption film, the adsorption film covers on the inlet of nucleic acid extraction column, but brokenly the part arrange the inlet of nucleic acid extraction column with between the adsorption film, and fix on the inner wall of nucleic acid purification post, can break and be provided with the recess on the part, the bottom of mixing board is for being used for disclosing brokenly the conical structure of recess, the cover is equipped with the mixing piece on the mixing board, the external diameter of mixing piece is less than the internal diameter of nucleic acid extraction column.
Further, the nucleic acid extraction system further comprises: the nucleic acid extracting device comprises a first slide rail, a mixing plate, a liquid transferring device and a positive pressure device are arranged side by side, the first slide rail is arranged along the mixing plate, the liquid transferring device and the positive pressure device are arranged in the direction on the mounting frame, the first slide rail is correspondingly arranged below the mixing rod, the infusion apparatus and the plug, and the nucleic acid extracting plate is arranged on the first slide rail in a sliding manner.
Further, the nucleic acid extraction system further comprises: nucleic acid collecting plate, elevating gear and spacing card frame drive arrangement, spacing card frame slidable mounting is in on the first slide rail, elevating gear arranges in the below of first slide rail, elevating gear includes elevator motor, elevating gear lead screw, elevating gear screw, lift limiting plate, lifter plate and is used for sliding the second slide rail that supports the nucleic acid collecting plate, elevator motor fixes on the mounting bracket, the elevating gear lead screw with elevator motor's output shaft transmission is connected, the elevating gear screw cover is established on the elevating gear lead screw, the last lift draw-in groove of having seted up of lift limiting plate, the elevating gear board dress is in the lift draw-in groove, the elevating gear screw passes the lift limiting plate with the bottom surface fixed connection of lifter plate, the nucleic acid collecting plate arranges on the top surface of lifter plate, the second slide rail is flush with the top surface of the lifting limit plate and is arranged in parallel with the first slide rail,
spacing card frame drive arrangement includes: a belt, a belt pulley, a fixed rod and a motor fixed on the mounting rack, wherein the fixed rod is fixed on the side wall of the first slide rail, the number of the belt pulleys is two, the two belt pulleys are respectively arranged on the output shaft of the motor and the fixed rod, the belt is sleeved on the two belt pulleys, the belt is arranged in parallel with the first slide rail, the limit clamping frame is fixed on the belt, the top surface of the limiting clamping frame is provided with a first clamping groove for clamping the nucleic acid extracting plate, the nucleic acid extracting plate is arranged in the first clamping groove, the bottom surface of the limiting clamping frame is provided with a second clamping groove for clamping the nucleic acid collecting plate, before the nucleic acid is eluted, the nucleic acid collecting plate is clamped on the second clamping groove through the lifting device, and the nucleic acid collecting plate is provided with nucleic acid collecting pipes which correspond to the nucleic acid extracting columns one by one.
Specifically, the nucleic acid extraction system further comprises: the clamping frame and the clamping driving device are used for clamping the nucleic acid extracting plate, clamping claws are rotatably arranged at the bottom of the clamping frame corresponding to the edge of the nucleic acid extracting plate, return springs for returning the clamping claws are arranged between the clamping claws and the clamping frame, the clamping driving device is fixed on the mounting frame, and the clamping driving device is configured to drive the clamping frame to move up and down.
Further, the nucleic acid extraction system further comprises: the second partition plate is arranged below the clamping jaw, the second partition plate driving device is fixed on the mounting frame, and the second partition plate driving device is configured to drive the second partition plate to horizontally move below the clamping jaw.
Specifically, a liquid level detection device is mounted on the side wall of the positive pressure provider.
The technical scheme provided by the embodiment of the invention has the following beneficial effects: according to the nucleic acid extraction system provided by the embodiment of the invention, the transfusion condition of each transfusion unit is controlled through the reagent flow controller, when chemical reagent in a certain reagent supplier is required to be output, the chemical reagent can be conveyed into a sealed cavity of a transfusion device from the reagent supplier in a timed and quantitative mode through the reagent flow controller, and high-pressure output is realized through the needle head, so that the nucleic acid extraction system provided by the embodiment of the invention does not need a liquid transfer work station and a liquid transfer suction head, the liquid transfer speed and efficiency are further improved, and the cost is reduced. Meanwhile, the plug is communicated with the nucleic acid extraction column in a sealing way, and high-pressure gas is introduced into the nucleic acid extraction column by using the positive pressure provider, so that liquid in the nucleic acid extraction column can rapidly pass through and flow out, and the nucleic acid extraction efficiency is further improved.
Drawings
In order to more clearly illustrate the technical solutions in the embodiments of the present invention, the drawings needed to be used in the description of the embodiments will be briefly introduced below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and it is obvious for those skilled in the art to obtain other drawings based on these drawings without creative efforts.
FIG. 1 is a schematic diagram showing the structure of a nucleic acid extraction system according to an embodiment of the present invention;
FIG. 2 is a schematic structural diagram of an infusion set provided by an embodiment of the invention;
FIG. 3 is a schematic structural diagram of a positive pressure apparatus according to an embodiment of the present invention;
FIG. 4 is a schematic structural diagram of a first partition and a first partition driving device according to an embodiment of the present invention;
FIG. 5 is a schematic structural view of a mixing rod coupled to an adsorption column according to an embodiment of the present invention;
FIG. 6 is a schematic structural view of a mixing rod in combination with a filter column according to an embodiment of the present invention;
fig. 7 is a schematic structural diagram of a lifting device provided in an embodiment of the present invention;
fig. 8 is a schematic structural diagram of a limit card frame driving device and a limit card frame according to an embodiment of the present invention;
fig. 9 is a schematic structural diagram of an infusion set driving unit provided by the embodiment of the invention.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, embodiments of the present invention will be described in detail with reference to the accompanying drawings.
Examples
An embodiment of the present invention provides a nucleic acid extraction system, as shown in fig. 1, including: a mounting frame 13, a nucleic acid extracting plate 1, and a pipetting device and a positive pressure device 7 which are arranged side by side.
The liquid transfer device includes: transfusion system 3, transfusion system mounting bracket 3a, transfusion system drive unit 3b and a plurality of infusion unit, combine fig. 2, transfusion system 3 includes the sealed chamber 5 with a plurality of infusion unit one-to-ones, each sealed chamber 5 is installed together side by side, syringe needle 6 is all installed to the bottom of every sealed chamber 5, every syringe needle 6 communicates with the sealed chamber 5 that corresponds separately respectively, transfusion system 3 installs the bottom at transfusion system mounting bracket 3a, transfusion system drive unit 3b fixes on mounting bracket 13, transfusion system drive unit 3b is configured to reciprocate through transfusion system mounting bracket 3a drive transfusion system 3.
Each infusion unit comprises: a reagent supply 8 and a reagent flow controller 9, wherein for any one of the infusion units, the outlet of the reagent supply 8 communicates with the inlet of the reagent flow controller 9, and the outlet of the reagent flow controller 9 communicates with each of the seal chambers 5. When the reagent supplying device is used, a liquid outlet of the reagent supplying device 8 is communicated with a liquid inlet of the reagent flow controller 9 through a reagent output pipe, a liquid outlet of the reagent flow controller 9 is also communicated with each sealing cavity 5 through a reagent output pipe, and the reagent output pipe can be a rubber hose. The rubber hose has certain flexibility, can adapt to reagent supplier 8, reagent flow controller 9 and seal chamber 5 of different positions to can rationally arrange reagent supplier 8, reagent flow controller 9 and seal chamber 5, reduce the volume of nucleic acid extraction system. In this embodiment, 8 needles 6 are mounted at the bottom of each sealed chamber 5, and the 8 needles 6 are arranged side by side. A water level alarm (not shown) may be provided in the reagent supplier 8 to remind an operator of timely replenishing the reagent in the reagent supplier 8 when the reagent in the reagent supplier 8 is about to be used up.
As shown in fig. 3, the positive pressure device 7 includes: the air conditioner comprises a plug 10, a positive pressure provider 12 and a positive pressure driving device 7a, wherein a vent pipe communication hole is formed in the plug 10, an air outlet of the positive pressure provider 12 is in sealed communication with the plug 10 through the vent pipe communication hole, the positive pressure driving device 7a is fixed on a mounting frame 13, and the positive pressure driving device 7a is configured to drive the positive pressure provider 12 to move up and down. The pressure provided by the positive pressure provider 12 may be 100kPa or more. The positive pressure supply device 12 may be an air compression pump, a piston pump, a diaphragm pump, or a liquid nitrogen bottle. In the present embodiment, the positive pressure device 7 may be provided in plurality, and a plurality of positive pressure devices 7 are provided side by side.
Referring to fig. 1 again, a plurality of nucleic acid extraction columns 1a are arranged on the nucleic acid extraction plate 1, the plurality of nucleic acid extraction columns 1a are correspondingly arranged below the needle 6 or the plug 10, a sealing gasket 11 in sealing fit with the plug 10 is arranged on a liquid inlet of the plurality of nucleic acid extraction columns 1a, and the plurality of nucleic acid extraction columns 1a are detachably and hermetically connected with the plug 10. In this embodiment, the number of the nucleic acid extraction columns 1a may be 96, a 96-well plate may be used in implementation, the number of the plugs 10 may be multiple, each plug 10 is respectively communicated with the air outlet of the positive pressure provider 12 through the ventilation pipe communication hole, and the multiple plugs 10 may be arranged in one-to-one correspondence with the multiple nucleic acid extraction columns 1 a.
The working principle of the invention is briefly described as follows:
the 5-6 chemical reagents for extracting or purifying nucleic acid are transferred into different reagent supply devices 8 by manual operation. The chemical reagent flows from the liquid outlet of the reagent supplier 8 to the liquid inlet of the reagent flow controller 9, the chemical reagent can be conveyed into the sealed cavity 5 of the transfusion device 3 through the reagent flow controller 9, and the liquid outlet amount of the needle 6 on different sealed cavities 5 is controlled or the liquid outlet stopping operation is started through the plurality of reagent flow controllers 9 respectively. When the nucleic acid extraction plate 1 is moved to the lower part of the corresponding needle 6 to prepare for adding liquid into the nucleic acid extraction column 1a, the infusion apparatus driving unit 3b can drive the infusion apparatus 3 to move downwards to the upper part of the nucleic acid extraction column 1a for adding liquid, after the liquid adding is finished, the infusion apparatus driving unit 3b can drive the infusion apparatus 3 to move upwards again, so that the needle 6 is far away from the nucleic acid extraction column 1a, the subsequent positive pressure operation is carried out, the nucleic acid extraction plate 1 is moved to the lower part of the plug 10, the positive pressure supply 12 is driven to move downwards by the positive pressure driving device 7a, the plug 10 is further hermetically installed on the liquid inlet of the corresponding nucleic acid extraction column 1a, the positive pressure supply 12 is started, the positive pressure device 7 can provide high-pressure gas, and the high-pressure gas acts on the surface of a solution (the solution comprises a nucleic acid solution or a chemical reagent to, because the high pressure provided by the positive pressure device 7 directly acts on the surface of the solution in each nucleic acid extraction column 1a, the solution can rapidly pass through the nucleic acid extraction column 1a and flow out, and after the positive pressure is finished, the positive pressure driving device 7a drives the positive pressure provider 12 to move upwards, so that the plug 10 is separated from the liquid inlet of the nucleic acid extraction column 1 a.
In the pipetting process, when different chemical reagents need to be transferred in the same sealed cavity 5, a certain passage through which the chemical reagents flow can be clear and clean in advance, and the specific method can comprise the following steps: a large amount of purified water is filled into the reagent supply device 8 for many times, and is output at high pressure through the needle 6, so that the cleaning of the passage is realized in the process of outputting the large amount of purified water at high pressure.
Specifically, the nucleic acid extraction system may further include: mixing plate 14, mixing plate mounting bracket 14a and mixing plate drive arrangement 14b, mixing plate mounting bracket 14a is fixed on mounting bracket 13, mixing plate drive arrangement 14b drive is installed on mixing plate mounting bracket 14a, mixing plate 14 is installed in mixing plate mounting bracket 14 a's bottom, mixing plate 14's bottom is provided with the mixing stick 4 with a plurality of nucleic acid extraction post 1a one-to-one, mixing plate drive arrangement 14b is configured to reciprocate in nucleic acid extraction post 1a through mixing plate mounting bracket 14a drive mixing stick 4. When the nucleic acid extraction plate 1 moves below the mixing plate 14, the mixing plate 14 moves up and down, so that the solution in the nucleic acid extraction plate 1 can be mixed.
Further, as shown in fig. 1 and 4, the nucleic acid extraction system may further include: a first partition 17 and a first partition driving means 17a, the first partition 17 being disposed between the kneading rod 4 and the nucleic acid extracting plate 1, the first partition driving means 17a being fixed to the mounting frame 13, and the first partition driving means 17a being configured to drive the first partition 17 to move horizontally below the kneading rod 4. After the solution in will nucleic acid extraction board 1 is mixed through mixing board 14, a small amount of solution is probably remained on mixing rod 4, stretch out first baffle 17 through first baffle drive arrangement 17a, make the solution that drips on mixing rod 4 fall on first baffle 17, thereby prevent that nucleic acid extraction board 1 from moving the process, remaining solution drips on mixing rod 4 and falls other downtheholely of nucleic acid extraction board 1, avoid the pollution problem of nucleic acid extraction in-process, when needs carry out the mixing operation once more, then retract first baffle 17 through first baffle drive arrangement 17 a.
Further, fig. 5 is a schematic structural diagram of an adsorption column provided in the embodiment of the present invention. As shown in fig. 5, the nucleic acid extraction column 1a may include: the adsorption column, the adsorption column includes breakable part 19 and adsorption film 27, adsorption film 27 covers on the inlet of nucleic acid extraction column 1a, breakable part 19 is arranged between inlet and adsorption film 27 of nucleic acid extraction column 1a to fix on the inner wall of nucleic acid purification column 1a, breakable part 19 is last to be provided with recess 19a, the bottom of mixing rod 4 is the tapered structure that is used for stabbing recess 19a, the cover is equipped with mixing piece 16a on mixing rod 4, the external diameter of mixing piece 16a is less than the internal diameter of nucleic acid extraction column 1 a. In practice, the kneading pieces 16a may be provided at the lower part of the kneading bar 4. The groove 19a may include an annular groove disposed at the center of the breakable member 19 and a plurality of strip-shaped grooves disposed between the annular groove and the outer wall of the breakable member 19 and disposed along a radius of the breakable member 19. When extracting plasmid DNA, can use adsorption column. Meanwhile, the nucleic acid extraction column 1a is internally provided with a breakable part 19, when the breakable part 19 is not broken in the nucleic acid extraction column 1a, the nucleic acid extraction column 1a can be matched with the breakable part 19 to be used as a test tube (without a water outlet at the bottom) for carrying and retaining liquid, so that various chemical reagents can mutually react. After the breakable part 19 is broken in the nucleic acid extraction column 1a, the liquid can flow out through the breakable part 19, so that the function of the nucleic acid extraction and purification column can be achieved, and the situation that a test tube and the nucleic acid extraction and purification column are used in the nucleic acid extraction process can be avoided, and the effects of improving the liquid transfer speed and reducing the cost are achieved.
Further, fig. 6 is a schematic structural diagram of a filter column according to an embodiment of the present invention. As shown in FIG. 6, the nucleic acid extraction column 1 comprises: a filter column comprising a breakable member 19 and a plurality of filter elements 20. The breakable component 19 is arranged between the liquid inlet of the nucleic acid extraction column 1a and the liquid outlet of the nucleic acid extraction column 1a and fixed on the nucleic acid purification column 1a, the breakable component 19 is provided with a groove 19a, the bottom of the mixing rod 4 is of a conical structure for breaking the groove 19a, the mixing rod 4 is sleeved with a mixing sheet 16a, the outer diameter of the mixing sheet 16a is smaller than the inner diameter of the nucleic acid extraction column 1a, a plurality of filtering pieces 20 are arranged between the breakable component 19 and the liquid outlet of the nucleic acid extraction column 1a and respectively fixed on the inner wall of the nucleic acid extraction column 1a, each filtering piece 20 is provided with a plurality of filtering micropores, and in two adjacent filtering pieces 20, the pore diameter of each filtering micropore of the filtering piece 20 close to the liquid inlet of the filtering column is larger than the pore diameter of each filtering micropore of the filtering piece 20 close to the liquid outlet of the.
In the present embodiment, the pores of the plurality of filter pores of each filter member 20 are equal in size. Thus, the solution can be ensured to uniformly flow through the filtering micropores, and the filtering effect is ensured.
Further, the plurality of filter elements 20 may include: at least three of the first filtering piece 20a, the second filtering piece 20b, the third filtering piece 20c, the fourth filtering piece 20d and the fifth filtering piece 20e, a plurality of first filtering micropores are formed in the first filtering piece 20a, the pore diameters of the first filtering micropores are all 20-25 μm, a plurality of second filtering micropores are formed in the second filtering piece 20b, the pore diameters of the second filtering micropores are all 16-19 μm, a plurality of third filtering micropores are formed in the third filtering piece 20c, the pore diameters of the third filtering micropores are all 10-15 μm, a plurality of fourth filtering micropores are formed in the fourth filtering piece 20d, the pore diameters of the fourth filtering micropores are all 5-9 μm, a plurality of fifth filtering micropores are formed in the fifth filtering piece 20e, and the pore diameters of the fifth filtering micropores are all 1-4 μm. Set up the filtration piece 20 that a plurality of apertures are different, and the aperture can be adjusted according to the size of solid phase material and the size of plasmid in the solution of treating the filtration to guarantee that solid phase material homoenergetic is filtered, and the plasmid can flow out smoothly.
Further, the plurality of filter elements 20 includes: the first filter element 20a, the second filter element 20b, the third filter element 20c, the fourth filter element 20d and the fifth filter element 20e, the pore size of the first filter pores is 22 μm, the pore size of the second filter pores is 18 μm, the pore size of the third filter pores is 13 μm, the pore size of the fourth filter pores is 8 μm, and the pore size of the fifth filter pores is 2 μm. The filtering effect of the filtering column can be ensured by the arrangement.
Further, the filter column may further include a spacer ring 20f, and the spacer ring 20f is disposed between two adjacent filter members 20 and fixed to an inner wall of the filter column. The spacer ring 20f can maintain the distance between two adjacent filtering members 20, thereby preventing the two adjacent filtering members 20 from being stuck together to affect the filtering efficiency. In this embodiment, the spacer ring 20f may be a polypropylene spacer ring, a polyethylene spacer ring, or a polytetrafluoroethylene spacer ring.
The filtering member 20 can be a filtering sieve plate, a filtering membrane and filtering cotton, and the filtering sieve plate, the filtering membrane and the filtering cotton can be matched and applied in the same filtering column.
Referring again to fig. 3, a liquid level detection device 30 is mounted on a side wall of the positive pressure provider 12. When implemented, the liquid level detection device 30 may be a non-contact liquid level meter, and specifically may be an ultrasonic sensor or an infrared sensor. The liquid level height after the action of the positive pressure device 7 can be monitored through the liquid level detection device 30, when the breakable part 19 breaks, liquid flows to the upper surface of the filter element 20 through the breakable part 19, at this time, if large impurities (such as blood clots) are mixed in nucleic acid to be extracted and purified, the large substances may be blocked on the filter element 20 of the filter column, so that the liquid cannot smoothly pass through the filter element 20, a large amount of liquid is blocked at the upper surface of the filter element 20, so that the nucleic acid to be extracted cannot be completely extracted and purified, during specific operation, the nucleic acid extraction column 1a can be moved to the lower part of the liquid level detection device 30, the liquid level height in the nucleic acid extraction column 1a is obtained through the liquid level detection device 30, and whether the liquid completely passes through the filter element 20 is judged through the liquid level height. An alarm (not shown) may be disposed on the side wall of the positive pressure supplier 12, and the alarm is electrically connected to the liquid level detecting device 30, and can give an alarm signal if the liquid does not completely pass through the filtering member 20.
Specifically, the nucleic acid extraction system may further include: the first slide rail 21, the blending plate 14 and the liquid-transfering device and the positive pressure device 7 are arranged side by side, the first slide rail 21 is arranged on the mounting rack 13 along the arrangement direction of the blending plate 14, the liquid-transfering device and the positive pressure device 7, the first slide rail 21 is correspondingly arranged below the blending rod 4, the infusion apparatus 3 and the plug 10, and the nucleic acid extraction plate 1 is arranged on the first slide rail 21 in a sliding manner.
Further, fig. 7 is a schematic structural diagram of the lifting device according to the embodiment of the present invention. As shown in fig. 7, the nucleic acid extraction system may further include: a nucleic acid collecting plate 22, a lifting device 26, a limiting card frame 23 and a limiting card frame driving device. The limit clamp frame 23 is slidably mounted on the first slide rail 21. The lifting device 26 is arranged below the first slide rail 21, the lifting device 26 comprises a lifting motor 26a, a lifting device screw rod 26c, a lifting device screw nut 26e, a lifting limiting plate 26f, a lifting plate and a second slide rail 26g (see fig. 1) for slidably supporting the nucleic acid collecting plate 22, the lifting motor 26a is fixed on the mounting frame 13, the lifting device screw rod 26c is in transmission connection with an output shaft of the lifting motor 26a, the lifting device screw nut 26e is sleeved on the lifting device screw rod 26c, a lifting clamping groove 26d is formed in the lifting limiting plate 26f, the lifting plate is arranged in the lifting clamping groove 26d and moves along the lifting clamping groove 26d, the lifting device screw nut 26e penetrates through the lifting limiting plate 26f to be fixedly connected with the bottom surface of the lifting plate, the nucleic acid collecting plate 22 is arranged on the top surface of the lifting plate, the second slide rail 26g is flush with the top surface of the lifting limiting plate 26f and is, the lift plate is raised to the top face of the lift restriction plate 26f so that the height of the lift plate is the same as the height of the top face of the lift restriction plate 26f, so that the nucleic acid collection plate 22 can be moved by the lift plate onto the second slide rail 26g and slide on the second slide rail 26g following the stopper frame 23.
Fig. 8 is a schematic structural diagram of a limit card frame and a limit card frame driving device according to an embodiment of the present invention. As shown in fig. 8, the spacing clip frame driving device includes: the device comprises a belt 23a, belt pulleys 23b, fixing rods 23c and a motor 23d fixed on the mounting frame 13, wherein the fixing rods 23c are fixed on the side wall of the first slide rail 21, the number of the belt pulleys 23b is two, the two belt pulleys 23b are respectively installed on an output shaft of the motor 23d and the fixing rods 23c, the belt 23a is sleeved on the two belt pulleys 23b, the belt 23a is arranged in parallel with the first slide rail 21, a limiting clamping frame 23 is fixed on the belt 23a, a first clamping groove for clamping the nucleic acid extracting plate 1 is formed in the top surface of the limiting clamping frame 23, the nucleic acid extracting plate 1 is clamped in the first clamping groove, and a second clamping groove for clamping the nucleic acid collecting plate 22 is formed in the bottom surface of the limiting clamping frame. Before the nucleic acid is eluted, the nucleic acid collecting plate 22 is clamped on the second clamping groove by the ascending motion of the elevating device 26, and the nucleic acid collecting plate 22 is provided with nucleic acid collecting tubes 22a corresponding to the nucleic acid extracting columns 1a one by one (see fig. 1). In practice, a 96-well plate may be used as the nucleic acid collecting plate 22. When the nucleic acid collecting plate 22 is lifted, the lifting motor 26a is driven, the lifting motor 26a drives the lifting device screw rod 26c to rotate, the lifting device screw nut 26e moves up and down along the lifting device screw rod 26c, the lifting plate moves up and down in the lifting clamping groove 26d of the lifting limiting plate 26f along with the lifting device screw nut 26e, when the top surface of the lifting plate is flush with the top surface of the lifting limiting plate 26f, the nucleic acid collecting plate 22 can be clamped at the bottom of the limiting clamping frame 23, the nucleic acid collecting plate 22 can move along with the limiting clamping frame 23, at the moment, the nucleic acid collecting plate 22 moves to the second sliding rail 26g through the lifting plate along with the movement of the limiting clamping frame 23, and meanwhile, the nucleic acid collecting plate 22 moves to the designated position along with the limiting clamping frame 23.
In implementation, the number of the first slide rails 21 is two, the two first slide rails 21 are disposed on two sides of the limiting card frame 23, the number of the second slide rails 26g is also two, the two second slide rails 26g are disposed on two sides of the nucleic acid collecting plate 22, and the two second slide rails 26g are located between the two first slide rails 21.
Specifically, as shown in fig. 1, the nucleic acid extraction system may further include: a clamp frame 24 for clamping the nucleic acid extracting plate 1 and a clamp driving device 24a, wherein a jaw 24b is rotatably provided at the bottom of the clamp frame 24 corresponding to the edge of the nucleic acid extracting plate 1, a return spring 24c for returning the jaw 24b is provided between the jaw 24b and the clamp frame 24, the clamp driving device 24a is fixed to the mounting frame 13, and the clamp driving device 24a is configured to drive the clamp frame 24 to move up and down. Press from both sides and get drive arrangement 24a and can lift with pressing from both sides frame 24 whole, when pressing from both sides frame 24 whole decline, jack catch 24b through the corresponding nucleic acid extraction board 1 the border after, through reset spring 24c back that resets, the card is in the border bottom of nucleic acid extraction board 1, can avoid like this that the user is manual to take nucleic acid extraction board 1, has reduced user's work load on the one hand, and on the other hand has improved nucleic acid extraction system's degree of automation. In implementation, a horizontal shaft may be provided on the vertically arranged gripping frame 24, and the jaw 24b is mounted on the horizontal shaft, and at the same time, a bearing may be provided on the horizontal shaft, and the jaw 24b is mounted on the bearing, in order to facilitate rotation of the jaw 24 b.
When the nucleic acid extracting plate 1 is a 96-well plate, the number of the claws 24b can be 4, and the 4 claws 24b are respectively arranged corresponding to four sides of the nucleic acid extracting plate 1, so that the stability of clamping the nucleic acid extracting plate 1 can be ensured, and the nucleic acid extracting plate 1 clamped by the clamping can be prevented from falling off.
Further, the nucleic acid extraction system may further include: a second partition plate 25 and a second partition plate driving means 25a, the second partition plate 25 being disposed between the claw 24b and the nucleic acid extracting plate 1, the second partition plate driving means 25a being fixed on the mounting frame 13, and the second partition plate driving means 25a being configured to drive the second partition plate 25 to move horizontally below the claw 24 b. When the claws 24b clamp the nucleic acid extracting plate 1, the solution remaining on the nucleic acid extracting plate 1 may drip out, and the second partition plate 25 is extended by the second partition plate driving means 25a, and the extended second partition plate 25 can receive the solution dripping out of the nucleic acid extracting plate 1.
In the present embodiment, as shown in fig. 4, the structure of the second partition driving device 25a is the same as that of the first partition driving device 17a, and the present embodiment takes the structure of the first partition driving device 17a as an example, and a specific structure of the second partition driving device 17a is described, specifically, the first partition driving device 17a includes: baffle driving motor 17b, baffle driving mounting panel 17c, baffle axis of rotation 17d, baffle belt 17e and baffle installation pole 17f, baffle driving mounting panel 17c is fixed on mounting bracket 13, baffle driving motor 17b fixes on the bottom of baffle driving mounting panel 17c, and baffle driving motor 17 b's output shaft and baffle axis of rotation 17d all pass baffle driving mounting panel 17c, baffle belt 17e arranges on the top surface of baffle driving mounting panel 17c, and baffle belt 17e overlaps respectively the transmission and establishes on baffle driving motor 17 b's output shaft and baffle axis of rotation 17d, the one end of baffle installation pole 17f is installed on baffle belt 17e, the other end of baffle installation pole 17f is installed in the bottom of first baffle 17. In other embodiments, the second barrier drive 25a and the first barrier drive 17a may each be driven in other manners.
Specifically, the reagent flow controller 9 may be: peristaltic pump, diaphragm pump, metering pump, syringe pump, plunger pump, piston pump, or self-priming pump.
The infusion set driving unit 3b, the positive pressure driving device 7a, the blending plate driving device 14b and the clamping driving device 24a provided in the embodiment of the present invention may all be of the same driving structure, as shown in fig. 9, which is briefly described by taking the infusion set driving unit 3b as an example, and specifically, the infusion set driving unit 3b includes: the infusion apparatus driving device comprises an infusion apparatus driving motor 3c, an infusion apparatus driving mounting plate 3d, an infusion apparatus belt 3e, a screw rod 3f, a screw rod nut block 3g, a screw rod nut slide block 3h and a screw rod nut first slide rail 3i, wherein the infusion apparatus driving mounting plate 3d is fixed on a mounting frame 13, the infusion apparatus driving motor 3c is fixed on the bottom of the infusion apparatus driving mounting plate 3d, an output shaft of the infusion apparatus driving motor 3c and the screw rod 3f both penetrate through the infusion apparatus driving mounting plate 3d, the infusion apparatus belt 3e is arranged on the top surface of the infusion apparatus driving mounting plate 3d, the infusion apparatus belt 3e is respectively sleeved on the output shaft of the infusion apparatus driving motor 3c and the screw rod 3f in a transmission manner, the screw rod nut block 3g is matched with the screw rod 3f through threads, the screw rod nut block 3g is fixed on the mounting frame, the feed screw nut slider 3h is fixed on the feed screw nut block 3g, and the feed screw nut slider 3h is slidably disposed on the feed screw nut first slide rail 3 i.
The following briefly introduces the working principle of the nucleic acid extraction system provided by the embodiment of the present invention, which is as follows:
first, 6 kinds of chemical reagents for extracting or purifying the corresponding nucleic acids are manually transferred to different reagent supplies 8.
1. If the extraction or purification of plasmid DNA nucleic acid, firstly, manually loading the nucleic acid extracting plate 1 with the nucleic acid extracting column 1a as the filter column with plasmid DNA cell solution to be extracted or purified, and the nucleic acid extracting column 1a has a breakable part 19 for preventing the solution from flowing downwards, aligning the liquid outlet of the nucleic acid extracting plate 1 with the nucleic acid extracting column 1a as the filter column with the liquid inlet of the nucleic acid extracting plate 1 with the nucleic acid extracting column 1a as the adsorption column, placing the two superposed nucleic acid extracting plates 1 (the nucleic acid extracting column 1a of the nucleic acid extracting plate 1 positioned above is the filter column, and the nucleic acid extracting column 1a of the nucleic acid extracting plate 1 positioned below is the adsorption column) on the limit card frame 23 (the start position) and clamping on the limit card frame 23. At this time, the two nucleic acid extracting plates superposed together can move together following the stopper frame 23. In addition, the nucleic acid collecting plate 22 is manually placed on the lifting plate, and the subsequent process is automatically completed by the system without manual operation.
Move spacing card frame 23 to the below of moving liquid device, make every row of filter column in the nucleic acid mentions board 1 receive in proper order move liquid device's below of the first row of syringe needle output of below 1 kind of chemical reagent after move to mixing board 14, make mixing stick 4 stretch into in the filter column of one-to-one and up-and-down motion, thereby make the solution mixing in the filter column, move upward mixing stick 4 after the mixing and leave the inlet of filter column, first baffle 17 extends between mixing stick 4 and the inlet of filter column, block probably from the liquid that drips on mixing stick 4 through first baffle 17, thereby prevent the pollution.
Move spacing card frame 23 to the below of liquid-transfering device, make every row of every filter column receive the 2 nd kind of chemical reagent of second row of syringe needle output of liquid-transfering device in proper order and move to the below of mixing board 14, make mixing stick 4 stretch into in the filter column and move from top to bottom to make the solution in the filter column mix, the cell rupture after plasmid DNA cell solution that treats extraction or purification and 2 kinds of chemical reagent in the filter column at this moment react, plasmid DNA dissolves in liquid, and genomic DNA and protein then take place the denaturation reaction and fuse into the debris of flocculent solid phase. The mixer plate 14 is moved a little further down the filter column so that the lower end of the mixer rod 4 breaks the breakable part 19 in the filter column and the liquid in the filter column flows down through the break in the breakable part 19 and onto the filter element 20.
Move spacing card frame 23 to the below of malleation device 7 and accept the malleation, the solution that contains plasmid DNA in the filter column loops through the breach, a plurality of filter 20 and the liquid outlet of filter column flows to the adsorption column in the below, and solid-phase debris is filtered by a plurality of filter 20 in the filter column through the breach.
The limiting clamp frame 23 is moved to the lower part of the clamping device 24, the clamping device 24 clamps the upper nucleic acid extracting plate 1 (the nucleic acid extracting plate 1 with the nucleic acid extracting column 1a as the filter column) and then lifts the nucleic acid extracting plate 1, so that the nucleic acid extracting plate 1 is separated from the lower nucleic acid extracting plate 1 (the nucleic acid extracting plate 1 with the nucleic acid extracting column 1a as the adsorption column), the second partition plate 25 extends to the lower part of the upper nucleic acid extracting plate 1, and the second partition plate 25 blocks liquid which possibly drips from the liquid outlet of the filter column 1a, thereby preventing pollution.
The limiting clamp frame 23 (at this time, only one nucleic acid extraction plate 1 with the nucleic acid extraction column 1a as the adsorption column on the limiting clamp frame 23) is moved to the lower part of the positive pressure device to sequentially make the nucleic acid extraction column 1a receive positive pressure, the solution in the adsorption column flows out to the waste liquid basin 15 below the nucleic acid extraction plate 1 with the nucleic acid extraction column 1a as the adsorption column, and plasmid DNA in the solution is adsorbed by the adsorption film 27 in the adsorption column.
The limiting clamping frame 23 is moved to the lower part of the liquid-transfering device, so that the nucleic acid extraction column 1a sequentially receives the 3 rd chemical reagent (first cleaning solution) output by the third row of needles, then moves to the lower part of the positive pressure device 7 and receives positive pressure, and when the first cleaning solution passes through the adsorption membrane, plasmid DNA on the membrane is cleaned, and then the first cleaning solution flows out of the waste liquid basin 15 below the nucleic acid extraction plate 1. When realizing, waste liquid basin 15 arranges on the mounting bracket 13 between two second slide rails 26g, and waste liquid basin 15 also can be inlayed on mounting bracket 13, and the bottom of waste liquid basin 15 can also be connected with the waste liquid bottle through the hose, and the waste liquid of collecting in the waste liquid basin 15 can be through in the hose inflow waste liquid bottle, treats that the waste liquid in the waste liquid bottle is collected the back, can pour the waste liquid and continue to collect after.
The limiting clamp frame 23 is moved to the lower part of the liquid-transfering device, so that the nucleic acid extraction column 1a sequentially receives the 4 th to 5 th chemical reagents (second cleaning solution and third cleaning solution) output by the fourth row to the fifth row of needles, and the subsequent process is the same as the first cleaning solution.
The limiting clamp frame 23 is moved to the initial position, the lifting motor 26a drives the lifting device screw rod 26c to rotate, so that the lifting device screw nut 26e drives the lifting plate to lift, at the moment, the nucleic acid collecting plate 22 positioned on the lifting plate is clamped at the bottom of the limiting clamp frame 23, the nucleic acid collecting plate 22 can move on the first slide rail 26g of the lifting device together with the limiting clamp frame 23, at the moment, a liquid inlet of a nucleic acid collecting pipe 22a on the nucleic acid collecting plate 22 below the limiting clamp frame 23 corresponds to a liquid outlet of a nucleic acid extracting column 1a above the limiting clamp frame 23, the limiting clamp frame 23 moves to the lower part of the liquid transferring device, so that an adsorption column of the nucleic acid extracting plate 1 sequentially receives 6 th chemical reagent eluent output by a sixth row of needle heads, then moves to the lower part of the positive pressure device 7 together to receive positive pressure, plasmid DNA is separated from an adsorption film of the adsorption column along with the eluent and flows into, the position-limiting clamping frame 23 with the adsorption plate 1 and the nucleic acid collection plate 22 clamped thereon is moved to the start position, and the extraction or purification process of the plasmid DNA is completed.
2. If the nucleic acid to be extracted or purified is genomic DNA, viral DNA or RNA, a filter plate is not required, and the process is relatively simple. Firstly, a nucleic acid cell solution to be extracted or purified is manually filled into the nucleic acid extracting plate 1 with the nucleic acid extracting column 1a as an adsorption column, a breakable part 19 is arranged in the adsorption column and used for preventing liquid from flowing downwards, and the nucleic acid extracting plate 1 with the nucleic acid extracting column 1a as the adsorption column is clamped on a limiting clamp frame 23, so that the nucleic acid extracting plate 1 with the nucleic acid extracting column 1a as the adsorption column can move along with the limiting clamp frame 23. In addition, the nucleic acid collecting plate 22 is manually placed on the lifting plate, and the subsequent process is automatically completed by the system without manual operation.
Moving the limiting card frame 23 to the lower part of the liquid-transfering device to make the adsorption column of the nucleic acid extraction plate 1 receive the 1 st chemical reagent outputted by the first row of needles in sequence, then moving to the lower part of the mixing plate 14 to make the solution pass through the mixing rod 4 for mixing, moving the mixing plate 14 upwards to make the mixing rod 4 leave the liquid inlet of the adsorption column, extending out the first partition plate 17 to make the first partition plate 17 block between the liquid inlet of the mixing rod 4 and the adsorption column, moving the limiting card frame 23 to the lower part of the liquid-transfering device to make the adsorption column of the nucleic acid extraction plate 1 receive the 2 nd chemical reagent outputted by the second row of needles in sequence, moving the limiting card frame 23 to the lower part of the mixing plate 14 for mixing, at this time, the cells are broken, the nucleic acid is released into the solution, moving the mixing plate 14a little distance downwards in the filtering column to the breakable part 19 to be broken by the rod 4, moving the limiting card frame 23 to the lower part of the positive pressure device 7 to make the column of the nucleic, the liquid flows out through the adsorption film 27 and is received by the waste liquid basin 15, the nucleic acid is adsorbed by the adsorption film 27, the limit card frame 23 is moved to the position below the liquid-transfering device to enable the adsorption columns of the nucleic acid extraction plate 1 to respectively and sequentially receive the 3 rd to 5 th chemical reagents (cleaning liquid) output by the third row of needles to the fifth row of needles, the cleaning liquid is added at each time and then is moved to the position below the positive pressure device 7, the cleaning liquid cleans the nucleic acid in the adsorption film 27 and then flows out to the waste liquid basin 15, the limit card frame 23 is moved to the starting position, the lifting device lead screw 26c is driven by the lifting motor 26a to rotate, the lifting device lead screw 26e drives the lifting plate to lift up, the nucleic acid collection plate 22 on the lifting plate is clamped at the bottom of the limit card frame 23, the nucleic acid collection plate 22 can move on the first slide rail 26g of the lifting device along with the limit card frame 23, the limit card frame 23 is moved to the position below the liquid-transfering device to enable the columns of the nucleic acid extraction Liquid removal), the limiting clamping frame 23 is moved to the lower part of the positive pressure device 7 to receive positive pressure in sequence, DNA on the adsorption film 27 is eluted into the nucleic acid collecting plate 22, the limiting clamping frame 23 is moved to the initial position, and the extraction or purification process of nucleic acid is completed.
The nucleic acid extraction system provided by the embodiment of the invention has the advantages that when certain chemical reagents are required for nucleic acid extraction or purification, the chemical reagent can be conveyed from the reagent supplier to the sealed cavity of the transfusion device in a timed and quantitative mode through the reagent flow controller, and high-pressure output of the reagent is realized through the needle, which enables the nucleic acid extraction system provided by the embodiment to avoid using an automatic pipetting workstation and a high-quality pipette tip, and then guaranteed the speed of moving liquid and reduced the cost that moves liquid, in addition, through end cap and nucleic acid extraction post sealed intercommunication, utilize the malleation provider to let in high-pressure gas in the nucleic acid extraction post, make the liquid in the nucleic acid extraction post can flow out through the resistance of post inner membrance fast, avoided the troublesome poeration that uses centrifuge or negative pressure machine to bring and the waste of time to further improve the efficiency of nucleic acid extraction and reduced the cost of extracting.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents, improvements and the like that fall within the spirit and principle of the present invention are intended to be included therein.
Claims (6)
1. A nucleic acid extraction system, comprising: a mounting rack, a nucleic acid extracting plate, a liquid-transfering device and a positive pressure device which are arranged side by side,
the liquid transfer device includes: the infusion set comprises sealing cavities which correspond to the infusion units one by one, the sealing cavities are arranged side by side, a needle is arranged at the bottom of each sealing cavity, each needle is communicated with the corresponding sealing cavity, the infusion set is arranged at the bottom of the infusion set mounting frame, the infusion set driving unit is fixed on the mounting frame and is configured to drive the infusion set to move up and down through the infusion set mounting frame;
each of the infusion units comprises: a reagent supply and a reagent flow controller, wherein for any one of the infusion units, a liquid outlet of the reagent supply is communicated with a liquid inlet of the reagent flow controller, and a liquid outlet of the reagent flow controller is communicated with each sealed cavity;
the positive pressure device includes: the device comprises a plug, a vent pipe, a positive pressure provider and a positive pressure driving device, wherein the plug is provided with a vent pipe communication hole, one end of the vent pipe is in sealed communication with the vent pipe communication hole, the other end of the vent pipe is in sealed communication with an air outlet of the positive pressure provider, the positive pressure driving device is fixed on the mounting frame and is configured to drive the positive pressure provider to move up and down, the pressure provided by the positive pressure provider is more than 100kPa, and a liquid level detection device is arranged on the side wall of the positive pressure provider and is used for detecting the liquid level in a nucleic acid extraction column on the nucleic acid extraction plate;
the nucleic acid extraction plate is provided with a plurality of nucleic acid extraction columns, the nucleic acid extraction columns are filter columns or adsorption columns, the nucleic acid extraction columns are correspondingly arranged below the needle heads or the plugs, a sealing gasket in sealing fit with the plugs is arranged on a liquid inlet of each nucleic acid extraction column, and the nucleic acid extraction columns are detachably and hermetically connected with the corresponding plugs;
the nucleic acid extraction system further comprises: the nucleic acid extraction column comprises a mixing plate, a mixing plate mounting rack and a mixing plate driving device, wherein the mixing plate mounting rack is fixed on the mounting rack, the mixing plate driving device is installed on the mixing plate mounting rack in a driving mode, the mixing plate is installed at the bottom of the mixing plate mounting rack, mixing rods in one-to-one correspondence with the nucleic acid extraction columns are arranged at the bottom of the mixing plate, and the mixing plate driving device is configured to drive the mixing rods to move up and down in the nucleic acid extraction columns through the mixing plate mounting rack;
the nucleic acid extraction system further comprises: the nucleic acid extracting plate is arranged on the first slide rail in a sliding manner;
the nucleic acid extraction system further comprises: the limiting clamping frame is slidably mounted on the first sliding rail, a liquid outlet of the nucleic acid extracting plate with the nucleic acid extracting column as a filter column is aligned with a liquid inlet of the nucleic acid extracting plate with the nucleic acid extracting column as an adsorption column, and the limiting clamping frame and the nucleic acid extracting plate are stacked together and placed on the limiting clamping frame;
the nucleic acid extraction system further comprises: the clamping frame and the clamping driving device are used for clamping the nucleic acid extracting plate, clamping claws are rotatably arranged at the bottom of the clamping frame corresponding to the edge of the nucleic acid extracting plate, return springs for returning the clamping claws are arranged between the clamping claws and the clamping frame, the clamping driving device is fixed on the mounting frame, and the clamping driving device is configured to drive the clamping frame to move up and down.
2. The nucleic acid extraction system according to claim 1, further comprising: the first partition plate is arranged between the mixing rod and the nucleic acid extraction plate, the first partition plate driving device is fixed on the mounting frame, and the first partition plate driving device is configured to drive the first partition plate to horizontally move below the mixing rod.
3. The nucleic acid extraction system of claim 1, wherein the nucleic acid extraction column comprises: a filter column comprising a breakable member and a plurality of filter members, the breakable member being disposed between a liquid inlet of the nucleic acid extraction column and a liquid outlet of the nucleic acid extraction column, and is fixed on the nucleic acid purification column, a groove is arranged on the breakable part, the bottom of the mixing rod is a conical structure for poking the groove, the mixing rod is sleeved with a mixing sheet, the outer diameter of the mixing sheet is smaller than the inner diameter of the nucleic acid extraction column, the plurality of filter members are disposed between the breakable member and the liquid outlet of the nucleic acid extraction column, and are respectively fixed on the inner wall of the nucleic acid extraction column, each filter element is provided with a plurality of filter micropores, and two adjacent filter elements are arranged in the filter element, the pore diameter of the filtering micropores of the filtering piece close to the liquid inlet of the filtering column is larger than that of the filtering micropores of the filtering piece close to the liquid outlet of the filtering column.
4. The nucleic acid extraction system of claim 1, wherein the nucleic acid extraction column comprises: adsorption column, adsorption column is including brokenly the part and adsorption film, the adsorption film covers on the liquid outlet of nucleic acid extraction column, brokenly the part and arrange the inlet of nucleic acid extraction column with between the adsorption film, and fix on the inner wall of nucleic acid purification post, can break and be provided with the recess on the part, the bottom of mixing board is for being used for disclosing brokenly the conical structure of recess, the cover is equipped with the mixing piece on the mixing board, the external diameter of mixing piece is less than the internal diameter of nucleic acid extraction column.
5. The nucleic acid extraction system according to claim 1, further comprising: nucleic acid collecting plate, elevating gear and spacing card frame drive arrangement, spacing card frame slidable mounting is in on the first slide rail, elevating gear arranges in the below of first slide rail, elevating gear includes elevator motor, elevating gear lead screw, elevating gear screw, lift limiting plate, lifter plate and is used for sliding the second slide rail that supports the nucleic acid collecting plate, elevator motor fixes on the mounting bracket, the elevating gear lead screw with elevator motor's output shaft transmission is connected, the elevating gear screw cover is established on the elevating gear lead screw, the last lift draw-in groove of having seted up of lift limiting plate, the elevating gear board dress is in the lift draw-in groove, the elevating gear screw passes the lift limiting plate with the bottom surface fixed connection of lifter plate, the nucleic acid collecting plate arranges on the top surface of lifter plate, the second slide rail is flush with the top surface of the lifting limit plate and is arranged in parallel with the first slide rail,
spacing card frame drive arrangement includes: a belt, a belt pulley, a fixed rod and a motor fixed on the mounting rack, wherein the fixed rod is fixed on the side wall of the first slide rail, the number of the belt pulleys is two, the two belt pulleys are respectively arranged on the output shaft of the motor and the fixed rod, the belt is sleeved on the two belt pulleys, the belt is arranged in parallel with the first slide rail, the limit clamping frame is fixed on the belt, the top surface of the limiting clamping frame is provided with a first clamping groove for clamping the nucleic acid extracting plate, the nucleic acid extracting plate is arranged in the first clamping groove, the bottom surface of the limiting clamping frame is provided with a second clamping groove for clamping the nucleic acid collecting plate, before the nucleic acid is eluted, the nucleic acid collecting plate is clamped on the second clamping groove through the lifting device, and the nucleic acid collecting plate is provided with nucleic acid collecting pipes which correspond to the nucleic acid extracting columns one by one.
6. The nucleic acid extraction system according to any one of claims 1 to 5, further comprising: the second partition plate is arranged below the clamping jaw, the second partition plate driving device is fixed on the mounting frame, and the second partition plate driving device is configured to drive the second partition plate to horizontally move below the clamping jaw.
Priority Applications (5)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810206072.XA CN110272808B (en) | 2018-03-13 | 2018-03-13 | Nucleic acid extraction system |
| US16/971,665 US11679382B2 (en) | 2018-03-13 | 2019-03-08 | Nucleic acid extraction system |
| EP19767872.5A EP3766955B1 (en) | 2018-03-13 | 2019-03-08 | Nucleic acid extraction system |
| PCT/CN2019/077403 WO2019174524A1 (en) | 2018-03-13 | 2019-03-08 | Nucleic acid extraction system |
| JP2020544535A JP7038439B2 (en) | 2018-03-13 | 2019-03-08 | Nucleic acid extraction system |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810206072.XA CN110272808B (en) | 2018-03-13 | 2018-03-13 | Nucleic acid extraction system |
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| Publication Number | Publication Date |
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| CN110272808A CN110272808A (en) | 2019-09-24 |
| CN110272808B true CN110272808B (en) | 2020-10-16 |
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| CN201810206072.XA Active CN110272808B (en) | 2018-03-13 | 2018-03-13 | Nucleic acid extraction system |
Country Status (5)
| Country | Link |
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| US (1) | US11679382B2 (en) |
| EP (1) | EP3766955B1 (en) |
| JP (1) | JP7038439B2 (en) |
| CN (1) | CN110272808B (en) |
| WO (1) | WO2019174524A1 (en) |
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2019
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- 2019-03-08 EP EP19767872.5A patent/EP3766955B1/en active Active
- 2019-03-08 JP JP2020544535A patent/JP7038439B2/en active Active
- 2019-03-08 US US16/971,665 patent/US11679382B2/en active Active
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| EP3766955A1 (en) | 2021-01-20 |
| US11679382B2 (en) | 2023-06-20 |
| CN110272808A (en) | 2019-09-24 |
| JP2021514629A (en) | 2021-06-17 |
| EP3766955B1 (en) | 2023-11-08 |
| EP3766955A4 (en) | 2022-01-05 |
| JP7038439B2 (en) | 2022-03-18 |
| US20200391198A1 (en) | 2020-12-17 |
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